In this study, we investigate the amebicidal activities of the
pharmaceutical triazole CYP51 inhibitors fluconazole,
itraconazole, and
voriconazole against Acanthamoeba castellanii and Acanthamoeba polyphaga and assess their potential as therapeutic agents against
Acanthamoeba infections in humans. Amebicidal activities of the
triazoles were assessed by in vitro minimum inhibition concentration (MIC) determinations using trophozoites of A. castellanii and A. polyphaga. In addition,
triazole effectiveness was assessed by
ligand binding studies and inhibition of CYP51 activity of purified A. castellanii CYP51 (AcCYP51) that was heterologously expressed in Escherichia coli.
Itraconazole and
voriconazole bound tightly to AcCYP51 (dissociation constant [Kd] of 10 and 13 nM), whereas
fluconazole bound weakly (Kd of 2,137 nM). Both
itraconazole and
voriconazole were confirmed to be strong inhibitors of AcCYP51 activity (50% inhibitory concentrations [IC50] of 0.23 and 0.39 μM), whereas inhibition by
fluconazole was weak (IC50, 30 μM). However,
itraconazole was 8- to 16-fold less effective (MIC, 16 mg/liter) at inhibiting A. polyphaga and A. castellanii cell proliferation than
voriconazole (MIC, 1 to 2 mg/liter), while
fluconazole did not inhibit Acanthamoeba cell division (MIC, >64 mg/liter) in vitro.
Voriconazole was an effective inhibitor of trophozoite proliferation for A. castellanii and A. polyphaga; therefore, it should be evaluated in trials versus
itraconazole for controlling
Acanthamoeba infections.